A preclinical study of allogeneic CD19 chimeric antigen receptor double‐negative T cells as an off‐the‐shelf immunotherapy drug against B‐cell malignancies

• Published in: Clinical & translational immunology Vol. 13; no. 12; pp. e70022 - n/a
• Main Authors: Wang, Dan, Wang, Liuyang, Liu, Shuai, Tong, Jianjun, Zhu, Honglin, Xu, Man, Li, Xiancai, Xiang, Zhiqiang, Sun, Qinghua, Wang, Hengcai, Wang, Yuli, Wang, Shuyang, Yang, Liming
• Format: Journal Article
• Language: English
• Published: Australia John Wiley & Sons, Inc 2024; John Wiley and Sons Inc; Wiley
• Subjects: allogeneic CD19‐CAR‐DNTs; Allografts; Antibodies; Antigens; Antitumor activity; Blood cancer; B‐cell malignancies; Cancer; CD19 antigen; Cell culture; Cell survival; Cell viability; Cells; Chimeric antigen receptors; Cryopreservation; Cytotoxicity; cytotoxicity and safety; Expression vectors; Flow cytometry; Genotype & phenotype; Graft versus host disease; Graft-versus-host reaction; Immunotherapy; Immunotoxicity; Ligands; Lymphocytes; Lymphocytes T; Lymphoma; Malignancy; Manufacturing; Nitrogen; off‐the‐shelf immunotherapy; Original; Proteins; Proteomes; Tumors; Xenografts; cytotoxicity and safety; B‐cell malignancies; off‐the‐shelf immunotherapy; allogeneic CD19‐CAR‐DNTs
• DOI: 10.1002/cti2.70022
• ISSN: 2050-0068

Objectives To evaluate the manufacturability, efficacy and safety of allogeneic CD19 chimeric antigen receptor double‐negative T cells (CD19‐CAR‐DNTs) as an off‐the‐shelf therapeutic cell product. Methods A membrane proteome array was used to assess the off‐target binding of CD19‐CAR. DNTs derived from healthy donors were transduced with lentiviral vectors encoding the CD19‐CAR. The manufacture of the CD19‐CAR‐DNTs was under GMP conditions, and their surface molecule expression patterns were characterised using flow cytometry. We investigated the off‐the‐shelf potential of CD19‐CAR‐DNTs by evaluating the cryopreserved CD19‐CAR‐DNTs in terms of cell viability as well as their cytotoxicity against various CD19+ target cell lines and primary patient blasts in vitro. We evaluated the persistence and safety of the cryopreserved CD19‐CAR‐DNTs in xenograft models in vivo. Results GMP‐grade CD19‐CAR‐DNTs were manufactured and cryopreserved for use in advance. The cryopreserved CD19‐CAR‐DNTs maintain their viability and antitumor activity against various CD19+ target cell lines and primary patient blasts. These cells significantly prolonged the survival in Raji‐Luc‐xenografted NOG mice. Multiple infusions of the cells can further augment their efficacy. Remarkably, following a single infusion in mice, CD19‐CAR‐DNTs rapidly got distributed among well‐perfused organs initially, and progressively spread to most tissues, peaking at Day 43. In toxicity studies, CD19‐CAR‐DNTs significantly reduced tumor burden and ameliorated tissue damage in tumor‐bearing NOG mice. Critically, no immunotoxicity or graft versus host disease was observed in non‐tumor‐bearing NOG mice. Conclusions The allogeneic CD19‐CAR‐DNTs fulfil the requirements of an off‐the‐shelf therapeutic cell product, offering a promising new approach to the treatment of haematological malignancies. In this study, we found that a novel off‐the‐shelf CD19 double‐negative T cell (CD19‐CAR‐DNT) therapy demonstrated significant antitumor activity and safety in vitro and in vivo. GMP‐grade CD19‐CAR‐DNTs maintained efficacy after cryopreservation and significantly extended survival in tumor models. No toxicity or graft versus host disease was observed, offering a new strategy for the treatment of haematologic malignancies.